Variation of mini- and microsatellite DNA repeats in parthenogenetic lizard Darevskia armeniaca as revealed by DNA fingerprinting analysis

Population and family samples of two morphological forms (mutant and normal with respect to dorsal color) of pathogenetic lizard Darevskia armeniaca were examined by means of DNA fingerprinting using M13 mini- and (GATA)n and (TCC)n microsatellite DNA markers. The morphological forms examined were characterized by clonally inherited, species-specific patterns of the DNA markers, which were different from the species-specific DNA fingerprints of the other parthenogenetic species of the genus Darevskia (D. dahli. D. unisexualis, and D. rostombekovi). The mean index of similarity (S) obtained for a sample of 36 individuals from three isolated populations using three types of DNA markers was 0.966. This was similar to the variability level observed in D. dahli (0.962) (P > 0.05), but higher than that in D. unisexualis (0.950) (P < 0.05) and D. rostombekovi (0.875) (P < 0.01). Inheritance of M13 minisatellite and (TCC)n microsatellite DNA markers in the F1 offspring of parthenogenetic lizards was examined. It was shown that variability and clonal diversity of the fingerprint phenotypes observed in the populations and families of D. armeniaca could be at least partly explained by RFLP mutations in microsatellite repeats.     

Characteristics of radish lines and hybrids from the spectrum of multiple molecular forms of enzymes

We characterized radish lines from a genetic collection on the basis of six enzyme systems, identified genes controlling these enzymes, and examined joint inheritance of some biochemical and morphological traits.     

Evaluation of the relevance of using RAPD-analysis for revealing the phylogenic connections between cultivars of drum wheat (T. durum Desf.)

A comparison of similarity indices between 64 durum wheat cultivars calculated using pedigree analysis and RAPD method showed a correspondence between these two approaches to estimation of genetic diversity. The associations between the results of RAPD clustering and coefficients of parentage (chi 2 test) and the coefficient of correlation between similarity matrices were statistically significant. However, the correlation was rather weak while pedigree analysis and RAPD method did not yield completely identical estimates of genetic diversity in the set of cultivars studied.     

Analysis of mutations in the RET proto-oncogene in patients with medullary thyroid tumor

The spectrum of mutations of the RET protooncogene was analyzed in Russian patients with inherited or sporadic medullary thyroid carcinoma (MTC). Four RET exons (11, 13, 15, and 16) were subjected to molecular analysis, and mutations were revealed and identified in 47.4% (9/19) patients with sporadic MTC. In total, six mutations (including three new ones) were observed. The most common mutation affected codon 918 to cause substitution of methionine with threonine and accounted for 31.6% alleles. Analysis of exons 11 and 16 revealed four mutations in patients with inherited multiple endocrine neoplasia type 2 (MEN 2). Mutations were found in each patient. Thyroidectomy was performed in four asymptomatic carriers of RET mutations from three MET 2A families (in two families, affected relatives had bilateral pheochromocytoma). In two patients, analysis of the surgery material revealed MTC microfoci in both lobes of the thyroid gland. The results provide the ground for constructing a bank of genetic information on Russian MTC patients with the clinically verified diagnosis.     

Multiple allelism of the net gene in Drosophila melanogaster and Drosophila simulans

The net gene mutations are known to cause abnormal pattern of veining in all wing regions except for the first posterior cells. In natural populations of Drosophila melanogaster, the net alleles were identified, which differ in phenotypic expression from standard mutations. The mutants net-extra-analis from a population Belokurikha-2000 have only a single additional vein in the third posterior cell. A line from Chernobyl-1986 population have another nontypical allele netCh86 and shows a lower degree of abnormalities than that usually observed. About 10% of these flies have an additional vein fragment in the first posterior cell. In both males and females of D. simulans population Tashkent-2001, which exhibit netST91 mutation, a net of additional veins is formed as a specific additional fragment in the first posterior cell. The pattern of veining conferred by alleles net-extra-analis and netCh86 is altered to a lesser extent; these alleles are dominant with respect to alleles net2-45 and netST91, which cause more abnormalities. The heterozygotes for alleles netST9 and netCh86 and for Df(2) net62 deletion have an additional fragment in the first posterior cell and show similarly strong deviations from normal wing vein pattern. The natural net alleles correspond, presumably, to different molecular gene defects involved into uncertain local interactions with numerous modifying factors and other genes that specify the wing vein pattern.     

Population-genetic structure of Chuvashia (from data on eight DNA loci in the nuclear genome)

Population-genetic study of indigenous populations representing three ethnic Chuvash group: highland (Cheboksarsk and Morgaush district), lowland (Kanash district) and mid-lowland (Marposad district). Eight polymorphic DNA loci of the nuclear genome (VNTR/PAH, STR/PAH, VNTR/ApoB, VNTR/DAT1, APF, VNTR/eNOS, IVS6aGATT, and KM.19/PstI) were examined in the population of each district. For each of the four population, we estimated the allele and genotype frequency distributions at each polymorphic system, heterozygosities HS and between-population differences FST. In the combined Chuvash sample, HS = 0.464 and FST = 0.006. Loci VNTR(DAT) and VNTR(ApoB) showed highest between-population differentiation (0.009 < or = FST < or = 0.012), and loci IVS6aGATT, APF, VNTR/eNOS, and D7S23 (KM.19), lowest differentiation (0.001 < or = FST < or = 0.003). Analysis of genetic distances revealed somewhat higher genetic similarity between the Cheboksarsk and Morgaush populations belonging to the highland Chuvash group, whereas the highland Chuvash population from the Marposad district, which belong to the mid-lowland group, was more distant from the former populations.     

Molecular evolution of satellite DNA CLsat in lizards of the Darevskia species (Sauria: Lacertidae): correlation with species diversity

The structure and evolution of a satellite DNA family was examined in lizards from the genus Darevskia (family Lacertidae). Comparison of tandem units of repeated DNA (satDNA), CLsat, in all species from the genus Darevskia has shown that their variability is largely based on single-nucleotide substitutions, which constitute about 50 diagnostic positions underlying classification of the family into three subfamilies. Maximum differences between the subfamilies reached 25%. At this level of tandem unit divergence between the subfamilies, no cross-hybridization between them was observed (at 65 degrees C). The individual variability of one subfamily within the species was on average 5% while the variability between species consensuses within a subfamily was 10%. The presence of highly conserved regions in all monomers and some features of their organization show that satellites of all Darevskia species belong to one satDNA family. The organization of unit sequences of satellites CLsat and Agi also detected by us in another lizard genus, Lacerts s. str. was compared. Similarity that was found between these satellites suggests their relatedness and common origin. A possible pathway of evolution of these two satDNA families is proposed. The distribution and content of CLsat repeat subfamilies in all species of the genus was examined by Southern blotting hybridization. Seven species had mainly CLsatI (83 to 96%); three species, approximately equal amounts of CLsatI and CLsatIII (the admixture of CLsatII was 2-3%); and five species, a combination of all three subfamilies in highly varying proportions. Based on these results as well as on zoogeographic views on phylogeny and taxonomy of the Darevskia species, hypotheses on the evolution of molecular-genetic relationships within this genus are advanced.     

Comparative evaluation of the efficiency of the effect of very high frequency electromagnetic waves on platelet functional activity

A comparative analysis was made of the effect of two kinds of EMI MMD-radiation: EMI MMD-waves, generated by a vehicle "Jav-1 M" (42.2 and 53.5 HHz), and EMI MMD-waves exerting influence with frequencies of molecular spectrum of radiation and nitric oxide absorption (150.176-150.644 HHz), obtained with a specially created generator, with respect to their influence on the functional ability of platelets of unstable angina pectoris patients. It was shown that in vitro EMI MMD-fluctuations with frequencies of molecular spectrum of radiation and nitric oxide absorption exert a stronger inhibiting influence on the functional activity of platelets of unstable angina pectoris patients. Features of the action of various kinds of EMI MMD-effect on the activative-high-speed characteristics of platelet aggregation are shown.     

Cell cycle and formation of active form of oxygen in rodent fibroblasts

Changes in the levels of mRNAs encoding ion transporters (ATP1B1, NHE1, NKCC1), beta-actin, GAPDH, regulators of proliferation and apoptosis (p53, Bcl-2) and kinase hSGK, involved in cell water regulation, were studied using RT PCR in the peripheral human lymphocytes activated with phytohemagglutinin for 4-24 h. The common, "grouped", effect that was found was an increase in the levels of the studied mRNAs after an 8 h activation, sometimes preceded by a delay or slight decrease at the initial stage of 0-4 h. Apart from the common features, some differences were observed in the time courses and amplitudes of the responses of individual mRNAs. The arrangement of the individual mRNA responses in lymphocytes from different donors could differ significantly, thus indicating differential regulation of the studied mRNAs apart from the "grouped" effect. The data obtained confirmed our suggestion that regulation of ion transport at the level of mRNA could be involved in the changes of ion balance at the late stage of lymphocyte activation.